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Shroomy_Mclovin
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Registered: 11/23/13
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Questionable mycelium
#23835895 - 11/15/16 03:45 PM (7 years, 2 months ago) |
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I did 4 large jars and 2 halfpint jars of WBS and during PC i ran out of water after an hour and a half or so. Stupid mistake, i know but after afew contaminated runs, i wanted to sterilize good this one. so i took my chances and added hot water, and wound up cracking every jar, they didnt shadder but they definitely cracked the bottoms real good. So i went and ran with it, didnt want to make more jars. Next day i nocked them up with"creeper" spores from a syringe... well only one has any kind of growth, oddly it is the one with what i would consider too much moisture. The other jars dont seem to have any mycelium, mold or bacteria, but this jar has questionable groth that looks more like cotton. Solid white fluffy growth. Not straky and not your typical dense mycelium... any opinions? Thanks in advance
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ComebackKid
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I didn't read your post yet but from what I can tell from pics you have mold
When I did my first PC run I ran out of water due to pcing at waaaaay to high of a temp. I did the same thing you did and cracked one of my jars.
I did a search and RR said that contaminants can still make it through the cracked glass.
I took my chances as well and wrapped cling film around the jar and taped it up with packing tape. I ended up with clean spaw. Mind you, I was also inoculating with agar.
In your case I'm going to assume contamination came in through the cracks in your glass.
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Substrate surface conditions / Monotub prep and care
Look around you... Everything you see exists inside the mind. Consciousness, the awareness that is experiencing this mind, is peering in from outside the universe. Our individual experiences are all part of the universe's experience of itself
Edited by ComebackKid (11/15/16 03:56 PM)
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Tira



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That does not look like something you'd like to have in your jars. Contam for sure.
Next time don't take any risks and start over when accidents happen. It consumes more time to try fixing problems the accident causes, and mostly they can't be fixed. You don't want to cut corners. Doing a proper start is the way to go.
-------------------- Useful Links for Beginners The Basics AMU Teks Frank''s Teks Agar Noob Forum Reccomended Teks Agar for guaranteed spawn, Proper pasteurization for guaranteed substrate.
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ComebackKid
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Re: Questionable mycelium [Re: Tira]
#23836144 - 11/15/16 04:39 PM (7 years, 2 months ago) |
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Even though I still ended up with clean spawn it does NOT not rule out the luck factor by any means. I do not reccomend taping cling film to your jars, but rather reccomend PCing at a lowe temperature.
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Substrate surface conditions / Monotub prep and care
Look around you... Everything you see exists inside the mind. Consciousness, the awareness that is experiencing this mind, is peering in from outside the universe. Our individual experiences are all part of the universe's experience of itself
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Shroomy_Mclovin
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Registered: 11/23/13
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Lame-o.... well im screwed, i dont have any more syringes. I might pick up a kit from planet k to save some time(on shipping), then once i get some stong mycelium i will just expand into some clean PCed WBS...
Any experience with the PK kits?
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Shroomy_Mclovin
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Just remembered that i had 2(cc or mg i forget the measurement but it is filled to the 2 on the syringe) that was leftover from inoculation. I just threw the syringe in a ziplock and stored it in my fridge, contams can definitely be an issue. But from what i understand, peitri cultivation can be a good option because i can isolate good micelium. Any advice? I also ready that potato dextrose can be a better option for tissue culture, while Malt dextrose would be better for spores... can anyone give me some 411 on the method i should look into?
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spore-ty



Registered: 01/21/16
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Look up pasty whyte agar tek I use extra light malt extract and agar for the most part i really reccommend petri dishes so you can see your mycelium
Also from time to time when working with the same culture in on going transfers I might switch up my agar recipe with pda or something but your gonna ahave to look into a pour agar tek which is the same thing as the pasty tek
Just pour your agar in a sab or in front of a fh is the difference really
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mrmazdarx9
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Quote:
Shroomy_Mclovin said: Lame-o.... well im screwed, i dont have any more syringes. I might pick up a kit from VENDOR to save some time(on shipping), then once i get some stong mycelium i will just expand into some clean PCed WBS...
Any experience with the VENDOR?
Dont post vendors
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Shroomy_Mclovin
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Re: Questionable mycelium [Re: spore-ty]
#23839109 - 11/16/16 03:07 PM (7 years, 2 months ago) |
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Quote:
spore-ty said: Look up pasty whyte agar tek I use extra light malt extract and agar for the most part i really reccommend petri dishes so you can see your mycelium
Also from time to time when working with the same culture in on going transfers I might switch up my agar recipe with pda or something but your gonna ahave to look into a pour agar tek which is the same thing as the pasty tek
Just pour your agar in a sab or in front of a fh is the difference really
What kind of agar? Im outfront of the science store right now lol... gonna check out those teks you mentioned.. thanks so much for your assistance...
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The Mycologist
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It doesnt matter, Potato Dextrose/Malt Extract
-------------------- "That you are here—that life exists, and identity; That the powerful play goes on, and you will contribute a verse.” ― Walt Whitman, Leaves of Grass

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Shroomy_Mclovin
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Re: Questionable mycelium [Re: mrmazdarx9]
#23839121 - 11/16/16 03:10 PM (7 years, 2 months ago) |
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Quote:
mrmazdarx9 said:
Quote:
Shroomy_Mclovin said: Lame-o.... well im screwed, i dont have any more syringes. I might pick up a kit from VENDOR to save some time(on shipping), then once i get some stong mycelium i will just expand into some clean PCed WBS...
Any experience with the VENDOR?
Dont post vendors 
My bad did that absent mindedly bc i was thinking more on the lines of venders being spore/suppliers via internet.. rather than local shops, i see plenty of people recomending local suppliers and i never thought of it... however i thank you for your post which contributed NOTHING to my initial question.. and i say that with the upmost sarcasm
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Shroomy_Mclovin
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Quote:
The Mycologist said: It doesnt matter, Potato Dextrose/Malt Extract
Awesome... thanks... is there anything else that i MUST GET FROM SCIENCE store while im here or will these teks require things which i can get from regular stores/walmart... sorry to ask i just want to be quick im in town and i am about to hit traffic if i dobt head home soon.. if anything else is needed i need to get it asap. I already got malt dextros and peitri dishes
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The Mycologist
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Scalpel maybe
-------------------- "That you are here—that life exists, and identity; That the powerful play goes on, and you will contribute a verse.” ― Walt Whitman, Leaves of Grass

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ComebackKid
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Inoc loop maybe
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Substrate surface conditions / Monotub prep and care
Look around you... Everything you see exists inside the mind. Consciousness, the awareness that is experiencing this mind, is peering in from outside the universe. Our individual experiences are all part of the universe's experience of itself
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Shroomy_Mclovin
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You fkin dudes ROCK! Got my pietri dishes and malt extract agar... gonna start my research at home.. im a little confused though cas i glanced at that pasteywhite tek and it seems i need agar agar, whatever that is? Lol im hoping i dont need to turn around? Can i be recomended a tek which will work with nothing else from the sci store, aside from my malt extract agar and pietris... this is a little overwelming... like the first time i grew BRF or WBS. I know it will be simple once i do it cas im familiar with that situation... i just need to put my big boy pants on and maybe someone to hold my hand the first time (lol)
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ComebackKid
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Your MEA is a premix of malt extract and agar agar powder. Pasty tek gives you a recipe to make potato dextrose agar from scratch. You could make MEA from scratch too if you wanted. But with what you have now all you need to do is add water.
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Substrate surface conditions / Monotub prep and care
Look around you... Everything you see exists inside the mind. Consciousness, the awareness that is experiencing this mind, is peering in from outside the universe. Our individual experiences are all part of the universe's experience of itself
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Shroomy_Mclovin
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Hell yeah buddy! Can you recommend a retard-proof tek for MEA? I think this would be the best option for me rather than any mixing aside from water. Gotta keep it simple or might overwork my fragile brain at this point lol
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Shroomy_Mclovin
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All my searching is piinting toward creating MEA simular to what you explained with pasteywhites tek... but since im one step ahead as i purchased MEA, what's next? Im assuming i mix the MEA powder with water, boil, put in bottle, PC, then inside a SAB pour the PCed MEA directly into pietris, cool, then inoculate with spores? I cant find anything to back this up so yea a tek would be beautiful for my current supplies. Thanks a bunch
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ComebackKid
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--------------------
Substrate surface conditions / Monotub prep and care
Look around you... Everything you see exists inside the mind. Consciousness, the awareness that is experiencing this mind, is peering in from outside the universe. Our individual experiences are all part of the universe's experience of itself
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Shroomy_Mclovin
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Ugh i suck! So that basically tells me what ratios to mix. But a step by step tek would make me comfortable. I just dont know, do i put one drop from my spore syringe, were my steps i previously mentioned correct? Sorry i keep asking questions im just afraid of ruining the last lil bit of spores i have on hand...
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ComebackKid
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I believe the steps you mentioned to be correct. Make like 20ish plates up and put a single drop of solution in the center of maybe 6 plates after the agar solidifies. You want to wrap your plates with cling film I believe to keep contams out. Then wait for growth. I'm not 100% certain as I hAve only ever fucked with no pour agar.
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Substrate surface conditions / Monotub prep and care
Look around you... Everything you see exists inside the mind. Consciousness, the awareness that is experiencing this mind, is peering in from outside the universe. Our individual experiences are all part of the universe's experience of itself
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Shroomy_Mclovin
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Once again, thanks for your time...
For someone who is familiar with pour agar, does this tek explain my steps properly for MAE after i mix the 25g - 500ml solution?
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The Mycologist
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-------------------- "That you are here—that life exists, and identity; That the powerful play goes on, and you will contribute a verse.” ― Walt Whitman, Leaves of Grass

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Shroomy_Mclovin
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Registered: 11/23/13
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Thanks for all the help!whats the best option for sterilizing things that cant be flame sterilized, like SAB, peitris, spore syringe ect? Alcohol, bleach, peroxide or a combination? Once i pour the PCd water/MEA, while it solidifies, what do i do to keep contams away before i noc it up? Just leave everything inside SAB till it hardens? Do i wait for room temp or what?
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Shroomy_Mclovin
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Also, ivalready started, at this time i am PC the 25g(scaled) to 500ml... While i mixed the stuff in before shutting the lid, i got a spoon full of the WEA and let it cool outside the PC, so i had an idea how long it takes to harden. Well it is definitely room temp but the top is very watery while the bottom is a little gelly. Does it take time as well as temperature (cooling) to harden? I dont want to pour into my peitris and find out that it is too watery and i wasted my peitris... ugh this is frustrating? It is at or below room temp, and i digitally scalled to 25g and measured perfectly 500ml of water... is it just going to take time as well as cooling?
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Shroomy_Mclovin
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Any help with the watery situation? Been 2 hours, probably ready to start pouring... does this shit dry over time? I weighed 25g and perfectly measured 500ml. The stuff i spooned out prior to PC just seems too watery on top, im afraid of pouring in dishes and ruining due to water content It has been cool for almost 3 hours and it isnt actually watery, it has the consistency of snot, not as watery on top, but it has long cooled
Edited by Shroomy_Mclovin (11/17/16 06:58 PM)
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ComebackKid
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Yeah it will solidify once it cools down. It usually only takes 5 minutes or so for my no pour agar dishes to solidify after PC
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Substrate surface conditions / Monotub prep and care
Look around you... Everything you see exists inside the mind. Consciousness, the awareness that is experiencing this mind, is peering in from outside the universe. Our individual experiences are all part of the universe's experience of itself
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Shroomy_Mclovin
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Then my mixture cant be right, this shit is actually cold at this point.... does the 25g x 500ml ratio seem correct?
Like i said i pulled this stuff out of the jar i PC right before i closed the lid.. and that was 3 hours ago. It is on a spoon sitting on my counter, the stuff is cold at this point yet still feels like snot
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tombosley8
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Hey bud here is some good shit for agar and you should watch RR's vids lets grow mushroom series. http://www.mushroomvideos.com/Agar-Petri-Dishes also you should buy the vids they show the full videos and can really come in handy. Well worth it for all the info in them.
here are some more agar links and overall info for cleaning and isolating/cloning thanks to bodhisatta Agar links all of this can be done in a SAB just as easy as long as it is tall enough or your agar bottle isn't too tall.
Did you get parafilm at the science store to wrap your petris after inoculation?
And spitball Jedi's stuff is awesome all around SBJ's The Basics
Edited by tombosley8 (11/17/16 09:15 PM)
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Shroomy_Mclovin
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Re: Questionable mycelium [Re: tombosley8]
#23844289 - 11/18/16 06:45 AM (7 years, 2 months ago) |
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Badass post! Yea i saw a lot of those, but thanks for the post regardless 100% greatful. Its just trippy because as im understanding, by purchasing this Malt Extract agar powder i bypassed all the legwork of creating my own agar-mix. I have been researching a lot and I'm getting a better understanding as we all have, this is a big learning curve. I feel like i have the idea, but what im stuck on right now is, why is my agar so runny and what is my proper mix. I put the 25g x 500ml ratio in this batch of MAE and it is wayy too runny, i still have not opened my PC, but the test spoon is at the consistancy of strawberry-jelly. The label calls for 28mg x 1L(1000ml) which means i needed an extra gram of powder. But i dont think it should have been so runny though. Now im afraid to mess up another batch, i got the hook up from the science store because it was an order that was not picked up so the guy didnt want it to expire and sold me the $90 bottle for $30...
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Shroomy_Mclovin
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So i left it in the pc overnight and i finally opened the pc and took it out, knowing it was too cold to use but i was curious of the consistancy. It did harden to what i believe is the proper thickness. However it is very moist. It is like a solid-gel consistancy, but it feels like i just ran it under water.... so if agar is on the same basis of BRF/WBS than it is certainly too wet but it is a good solid.. however if it is more forgiving with moisture, like a LC than it would have probably worked perfectly.maybe a couple more G of powder would fix the excess moisture?
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LogicaL Chaos
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Is that agar?
Or jelly popsicle?
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Shroomy_Mclovin
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Registered: 11/23/13
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Lol not a popsicle but it does look a lil yummy... i pcd it in a mason jar cas i didn't have a good jar for pouring, but that particular jar had an inoculation plug that i was going to pull out to pour once i was ready to fill my pietri dishes.. so yeah i pulled that directly from the mason jar, it was a somewhat firm gel but seemed too wet... is excess moisture an issue with MEA?
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r.lutece
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Registered: 09/06/15
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That extra gram of powder might have been just the thing to soak up that extra moisture. It's a small amount, but just like in cooking, it can make a big difference.
-------------------- One goes into an experiment knowing one might fail. But one does not undertake an experiment knowing one HAS failed.
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tombosley8
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Re: Questionable mycelium [Re: r.lutece]
#23845181 - 11/18/16 12:45 PM (7 years, 2 months ago) |
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Slightly wet is not an issue IME. I'd rather that than too dry.
I have pooling water and pretty soft agar in most of my dishes and It has caused no problems.
The thing with adding more or less powder is you are altering the nute content since yours is premixed.
usually with slightly less nutes I get faster growth and more rhizo growth Which I prefer and then switch back for the plate that I plan to inoculate grains/etc with.
I like making my own mix for that reason, so I'm not altering the agar content when adjusting the nutes.
You can order the ingredients really cheap off amazon or ebay.
Just look for Light Malt Extract and agar agar powder. They are all pretty similar in consistency as I've used about 10 different brands.
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ComebackKid
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Throw that back in the PC and pour your plates with it. Looks perfect
--------------------
Substrate surface conditions / Monotub prep and care
Look around you... Everything you see exists inside the mind. Consciousness, the awareness that is experiencing this mind, is peering in from outside the universe. Our individual experiences are all part of the universe's experience of itself
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